Direct electrochemistry of cytochrome P450 27B1 in surfactant films |
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Authors: | Steve Y Rhieu Daniel R Ludwig Vince S Siu G Tayhas R Palmore |
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Institution: | aDivision of Engineering, Brown University, Providence, RI 02912, USA;bDivision of Biology and Medicine, Brown University, Providence, RI 02912, USA |
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Abstract: | We report the first direct electrochemistry of cytochrome P450 27B1 (CYP27B1) immobilized on an edge-plane pyrolytic graphite (EPG) electrode coated with a film of didodecyldimethylammonium bromide (DDAB). Cyclic voltammetry (CV) in a deoxygenated solution revealed excellent electrochemical reversibility with an average midpoint potential of −180 ± 5 mV vs. Ag/AgCl and an apparent surface coverage of (7.0 ± 2.5) × 1013 molecules per cm2. The rate of heterogeneous electron transfer between CYP27B1 and the EPG electrode was determined to be 3.5 ± 0.6 s−1. Upon addition of oxygen, a significant increase in cathodic current occurred, likely due to electrocatalytic reduction of dioxygen to peroxide and/or water by CYP27B1. Characterization of the electrochemical properties of CYP27B1 is an important first step toward developing a bioelectrochemical method for measuring vitamin D in serum. |
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Keywords: | Cytochrome P450 Direct electrochemistry Surfactant films Thin-film voltammetry 25-Hydroxyvitamin D3 |
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